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Anne-Sophie Delmarcelle Mylah Villacorte Anne-Christine Hick Christophe E. Pierreux 《Journal of visualized experiments : JoVE》2014,(88)
The thyroid is a bilobated endocrine gland localized at the base of the neck, producing the thyroid hormones T3, T4, and calcitonin. T3 and T4 are produced by differentiated thyrocytes, organized in closed spheres called follicles, while calcitonin is synthesized by C-cells, interspersed in between the follicles and a dense network of blood capillaries. Although adult thyroid architecture and functions have been extensively described and studied, the formation of the “angio-follicular” units, the distribution of C-cells in the parenchyma and the paracrine communications between epithelial and endothelial cells is far from being understood.This method describes the sequential steps of mouse embryonic thyroid anlagen dissection and its culture on semiporous filters or on microscopy plastic slides. Within a period of four days, this culture system faithfully recapitulates in vivo thyroid development. Indeed, (i) bilobation of the organ occurs (for e12.5 explants), (ii) thyrocytes precursors organize into follicles and polarize, (iii) thyrocytes and C-cells differentiate, and (iv) endothelial cells present in the microdissected tissue proliferate, migrate into the thyroid lobes, and closely associate with the epithelial cells, as they do in vivo.Thyroid tissues can be obtained from wild type, knockout or fluorescent transgenic embryos. Moreover, explants culture can be manipulated by addition of inhibitors, blocking antibodies, growth factors, or even cells or conditioned medium. Ex vivo development can be analyzed in real-time, or at any time of the culture by immunostaining and RT-qPCR.In conclusion, thyroid explant culture combined with downstream whole-mount or on sections imaging and gene expression profiling provides a powerful system for manipulating and studying morphogenetic and differentiation events of thyroid organogenesis. 相似文献
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Anne-Christine Hick Anne-Sophie Delmarcelle Mahé Bouquet Sabrina Klotz Tamara Copetti Celine Forez Patrick Van Der Smissen Pierre Sonveaux Jean-François Collet Olivier Feron Pierre J. Courtoy Christophe E. Pierreux 《Developmental biology》2013
The thyroid is a highly vascularized endocrine gland, displaying a characteristic epithelial organization in closed spheres, called follicles. Here we investigate how endothelial cells are recruited into the developing thyroid and if they control glandular organization as well as thyrocytes and C-cells differentiation. We show that endothelial cells closely surround, and then invade the expanding thyroid epithelial cell mass to become closely associated with nascent polarized follicles. This close and sustained endothelial:epithelial interaction depends on epithelial production of the angiogenic factor, Vascular Endothelial Growth Factor-A (VEGF-A), as its thyroid-specific genetic inactivation reduced the endothelial cell pool of the thyroid by >90%. Vegfa KO also displayed decreased C-cells differentiation and impaired organization of the epithelial cell mass into follicles. We developed an ex vivo model of thyroid explants that faithfully mimicks bilobation of the thyroid anlagen, endothelial and C-cells invasion, folliculogenesis and differentiation. Treatment of thyroid explants at e12.5 with a VEGFR2 inhibitor ablated the endothelial pool and reproduced ex vivo folliculogenesis defects observed in conditional Vegfa KO. In the absence of any blood supply, rescue by embryonic endothelial progenitor cells restored folliculogenesis, accelerated lumen expansion and stimulated calcitonin expression by C-cells. In conclusion, our data demonstrate that, in developing mouse thyroid, epithelial production of VEGF-A is necessary for endothelial cells recruitment and expansion. In turn, endothelial cells control epithelial reorganization in follicles and C-cells differentiation. 相似文献
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Mark AJ Roberts Elias August Abdullah Hamadeh Philip K Maini Patrick E McSharry Judith P Armitage Antonis Papachristodoulou 《BMC systems biology》2009,3(1):105-14
Background
Developing methods for understanding the connectivity of signalling pathways is a major challenge in biological research. For this purpose, mathematical models are routinely developed based on experimental observations, which also allow the prediction of the system behaviour under different experimental conditions. Often, however, the same experimental data can be represented by several competing network models. 相似文献7.
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S.A. Bartolotta A.S. Hick M.F. Rodriguez Brasco B. Mothes M. Campos 《Journal of experimental marine biology and ecology》2009,369(2):144-147
Marine sponges are a rich source of structurally and biologically active metabolites of biomedical importance. We screened polar and non-polar samples of crude extracts obtained from marine sponges collected in different locations of Argentinean south sea coast, as a novel approach for their characterization.The evaluation was performed using cytotoxic and genotoxic biomarkers such as mitotic index (MI), cell proliferation kinetics (CPK) and sister chromatid exchanges (SCE), monitored in vitro using peripheral blood lymphocytes. Statistical analysis was performed using two-way analysis of variance (ANOVA). The extracts evaluated belonged to: Callyspongia flabellata (BURTON, 1932) (Callyspongiidae); Plicatellopsis sp.(Suberitidae); Callyspongia fortis (RIDLEY, 1881) (Callyspongiidae); Clathria (Microciona) antarctica (TOPSENT, 1917) (Microcionidae); Spongia (Spongia) magellanica (THIELE, 1905) (Spongiidae); Halicnemia papillosa (THIELE, 1905) (Axinellidae); Cliona chilensis (THIELE, 1905) (Clionidae); Haliclona sp. 1; Haliclona sp. 2(Chalinidae).Genotoxicity studies revealed that the evaluated sponge extracts did not exhibit cytotoxic activity measured from mitotic index MI and cell proliferation kinetics(CPK). In contrast, sister chromatid exchanges (SCE) showed that the non-polar extract of Callyspongia fortis and the polar extract of Cliona chilensis presented significant differences in SCE frequency (p < 0.001), when compared with control cultures. These results emphasize the need to set up a standard battery of “in vitro” genotoxicity testing for new chemicals, pharmaceutical and drugs. 相似文献
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